OsRacD belonging to rice Rho family of the small GTP binding proteins,is a pivotal gene involved in rice photoperiod fertility conversion of photoperiod sensitive genic male sterile rice,which influences rice fertility via controlling the pollen tube growth.T20N site-directed mutation was introduced into its highly conserved G1 motif by PCR-mediated method to mimic its GDP-binding state based on the sequences alignment and conserved domains analysis.The prokaryotic expression vector of OsRacD and T20N-OsRacD were constructed,and the His6 tag fused proteins were expressed and purified from E.coli.After identified by Western blot,the GTP hydrolysis activities were detected.The results showed that the GTPase activities of T20N-OsRacD were significantly reduced comparing with that of OsRacD,suggested that OsRacD and T20N-OsRacD have different biochemical characteristics.

ObjectiveTo evaluate the effect of Yuxingchangzhi Decoction on level of norepinephrine (NE) and 5-hydroxyptamin (5-HT) in serum of patients with post-stroke depression. Methods76 patients with post-stroke depression were treated with Yuxingchangzhi Decoction or placebo for 4 weeks. Before and after treatment, all the patients were evaluated with the Hamilton Rating Scale for Depression (HAMD),while the level of NE and 5-HT in serum of patients were measured.ResultsThe total score of HAMD of the patients of Yuxingchangzhi Decoction group was significantly different before and after treatment (P<0.01),while that of the placebo group was not significantly different (P>0.05). The NE and 5-HT level in serum of patients of the Yuxingchangzhi Decoction group were significantly different before and after treatment (P<0.01),but that of the placebo group was not significantly different (P>0.05). ConclusionYuxingchangzhi decoction can raise NE,5-HT level in patients with post-stroke depression and alleviate symptoms of depression significantly.

Objective To explore the mechanism of therapeutic action of needle knife “regulating sinews and treating bones” on knee osteoarthritis (KOA) by observing the effect of needle knife intervention on KOA rabbit behaviors, mechanical characteristics of patellar ligament (PL), and expressions of interleukin-4 (IL-4), matrix metalloproteinase-3 (MMP-3) and aggrecan in knee cartilages.Methods Forty New Zealand rabbits were randomly allocated to blank, model, needle knife and electroacupuncture groups, 10 rabbits each. A rabbit model of KOA was made by the modified Videman method of immobilization in extension position. After model making, the needle knife and electroacupuncture groups received needle knife and electroacupuncture treatments, respectively. A behavioral assessment was made using the modified Lequesne MG knee grade evaluation method in every group after model making and treatment. The samples were taken after treatment. PL tension, stress relaxation and creep state were tested using a Bose Electro Force 3300 protracted test machine. Cartilage cell IL-4 expression was examined by ELISA. MMP-3 mRNA and aggrecan mRNA expressions were detected by real-time PCR.Results After model making, there was a statistically significant difference in the Lequesne MG score between the model and blank groups (P<0.01); there was no statistically significant difference in the Lequesne MG score between the needle knife or electroacupuncture group and the model group (P>0.05). There was a statistically significant post-treatment difference in the Lequesne MG score between the needle knife or electroacupuncture group and the model group (P<0.01,P<0.05) and between the needle knife and electroacupuncture groups (P<0.05). There were statistically significant post-treatment differences in PL maximum stress, maximum displacement, elastic modulus, stress relaxation rate and creep rate between the model and blank groups (P<0.01,P<0.05). There were statistically significant post-treatment post-treatment differences in PL maximum stress, maximum displacement, elastic modulus, stress relaxation rate and creep rate between the needle knife and model groups (P<0.01,P<0.05). There was a statistically significant post-treatment difference in elastic modulus between the electroacupuncture and model groups (P<0.01). The IL-4 content and aggrecan mRNA expression decreased significantly and MMP-3 mRNA expression increased significantly in the model group after model making and there were statistically significant differences compared with the blank group (P<0.01,P<0.05). After treatment, the IL-4 content increased significantly in the needle knife and electroacupuncture groups compared with the model group (P<0.01,P<0.05) and aggrecan mRNA expression tended to increase in the two groups. The regulation of aggrecan mRNA and MMP-3 mRNA expressions was better in the needle knife group than in the electroacupuncture groups, but there was no statistically significant difference compared with the model group (P>0.05).Conclusion The mechanism of action of needle knife treatment on KOA may be that it improves ligament mechanical characteristics, regulates intra-articular stress environment, and modulates aggrecan mRNA and MMP-3 mRNA expressions and inhibits cartilage degeneration through IL-4 mechanical signal pathway, to produce the therapeutic effect of “regulating sinews and treating bones”.

OBJECTIVE: To observe the effect of acupoint injection at "Yingxiang"(LI 20) and "Yintang"(GV 29) on nasal allergic reactions and the expression of histamine receptor H 1 and H 4 in nasal mucosa of allergic rhinitis (AR) rats, so as to reveal its mechanism underlying improvement of AR. METHODS: SD rats were randomized into normal, model, non-acupoint injection and acupoint injection groups (n=8 in each). The AR model was established by intraperitoneal (i.p.) injection of mixture of ovalbumin, aluminium hydroxide gel and normal saline (once every other day, for 7 times), and nasal drip of ovalbumin (on the following day of i.p.for 7 days). The mixture solution of lidocaine, dexamethasone (DXM) and transfer factor (0.1 mL/acupoint or non-acupoint) was injected into bilateral LI 20 and GV 29 in the acupoint injection group, or into the non-acupoints (about 5 cm below the armpit on both sides, and the middle point between the left "Houhai"[GV 1] and "Huantiao"[GB 30]), on the 1st , 5th, 9th, and 13th day after modeling. Symptoms of sneezing, nasal discharge, nose-rubbing, etc. were scored after the treatment. The expression levels of H1 R and H4 R proteins and genes in the nasal mucosa tissue were determined using immunohistochemistry and quantitative real-time PCR, respectively. RESULTS: Compared with the normal group, the symptom score and the expression levels of H1 R, H4 R proteins and genes in nasal mucosa were significantly increased in the model group (P<0.05). After acupoint injection for 4 times, the symptom score and the expression levels of H1 R, H4 R proteins and mRNAs were significantly down-regulated in the acupoint injection group relevant to the model group (P<0.05). There were no significant differences between the non-acupoint and the model groups in the abovementioned indexes (P>0.05), suggesting a specificity of the effect of acupoint injection. CONCLUSION: Acupoint injection can relieve the allergic symptoms of AR rats, which may be related to its effects in down-regulating the over expression of H1 R and H4 R proteins and genes in the nasal mucosa.

ObjectiveTo study the characteristic of apoptosis and the expression of interleukin-1β(IL-1β)and caspase-3, as well as the relation among them after spinal cord injury(SCI) in rats.MethodsAdult SD rats were divided into two groups randomly: sham group and spinal cord injury group. The rats were killed at 1 h, 8 h, 24 h, and 72h after surgery. The expression of IL-1β and caspase-3 was determined with immunohistochemistry and the apoptosis was labeled by TUNEL.ResultsThere was only a few cells that expressed low level IL-1β and caspase-3 in sham group, but they both enhanced in SCI group within 8 h after operation, and then decreased to the level similar to sham group 72 h after operation. The rates of TUNEL-positive cells were similar with that of IL-1β and caspase-3 after SCI. There was positive correlation among expression of IL-1β and caspase-3 and apoptosis index.ConclusionExpression of IL-1β and caspase-3 increase and a large number of TUNEL-positive cells present after SCI in rats, and there is positive correlation among them.

Objective To explore the relationship between fetal heart size and gestational weeks (GW). Methods The size of left atrium (LA), right atrium (RA), left ventricle (LV), right ventricle (RV), aorta (AO), pulmonary artery (PA), foramen ovale (FO), heart area (HA), thoracic area (TA), heart circumference (HC) and thoracic circumference (TC) were measured for 512 fetal hearts at 14-39 GW. The relationship between GW and the measurement was evaluated. Results The size of fetal heart chambers, AO, PA and ventricular septum (IVS) increased with the development of GW. The PA/AO, LA/RA, LV/RV, HC/TC and HA/TA were stable compared with different GW. Conclusion Fetal heart chambers increase with the development of GW. HA is correlated well with GW.

OBJECTIVE To study the changes in matrix metalloproteinase-9 (MMP-9) mRNA, tissue-inhibitor of metalloproteinase-1 (TIMP-1) mRNA and the ratio of MMP-9 mRNA/TIMP-1 mRNA in the lungs of paraquate poisoning rats, and to investigate the protective effects of sodium dimercaptopropane sulfonate (DMPS, Unithiol). METHODS One hundred and twenty male SD rats were divided into 12 groups (n=10) randomly: normal control, DMPS control, PQ poisoning 1, 3, 7, 14 and 28 d model groups, (DMPS+PQ) 1, 3, 7, 14 and 28 d groups. PQ poisoning model was established by intraperitoneal injecting paraquate. The expression of MMP-9 mRNA and TIMP-1 mRNA in lung tissues was detected by RT-PCR. RESULTS ①By observing the changes of action and observing the lung tissues sections, the rats' PQ poisoning models was successfully established. ②The histopathology of lung showed infiltration of inflammatory cell in acute phase(within 2 weeks), the inflammation decreased gradually after 2 weeks, hyperplasia of collagen and pulmonary fibrosis were instead. Howerer, the pathological changes were alleviated obviously in the (DMPS+PQ) groups. ③Compared with the PQ groups, the expression of MMP-9 mRNA and TIMP-1 mRNA in lungs diminished greatly in the DMPS+PQ groups after rats injected DMPS(P<0.05); the ratio of MMP-9/TIMP-1 mRNA was bigger at 7, 14 and 28 d in the DMPS+PQ groups after rats injected DMPS. CONCLUSION Down-regulation of the expression of MMP-9 mRNA and TIMP-1 mRNA and up-regulation ratio of MMP-9 mRNA/TIMP-1 mRNA by DMPS may be one of mechanisms by which pulmonary injury and pulmonary fibrosis are prevented in the acute paraquate poisoning.

Ac-Phe-Lys-PABC-DOX (PDOX) is a smart doxorubicin (DOX) prodrug designed to decrease toxicities while maintaining the potent anticancer effects of DOX. This study was aimed at elucidating the effectiveness and toxicities of DOX and PDOX in patient-derived MCF-7 breast cancer cells in vitro. The MCF-7 cells were exposed to both PDOX and DOX, and cytotoxicities, cell cycle and P53/P21 signaling alterations were studied. Abundant cathepsin B was found in the MCF-7 cells, and treatment with PDOX and DOX triggered dose- and time-dependent cytotoxicity and resulted in a significant reduction in cell viability. The IC50 of PDOX and DOX was 3.91 and 0.94 μmol/L, respectively. Both PDOX and DOX caused an up-regulation of the P53/P21-related signal pathway, and PDOX significantly increased expression of P53 and caspase 3, and arrested the cell cycle at the G1/G2 phase. As compared with DOX, PDOX reduced toxicities, and it may have different action mechanisms on breast cancer cells.

Objective To study the changes of serum level of high mobility group box-1(HMGB- 1)in patients with multiple trauma in order to forecast organ dysfunction(OD)and deaths.Methods The optical densities of HMGB-1 in serum of 35 patients with multiple trauma were determined on 1st,3rd, and 7th days after trauma,and the incidence of organ dysfunction and deaths were evaluated,then analyzed statistically to learn the relation between the serum levels of HMGB-1 and deaths with an attempt of predic- ting the incident of organ dysfunction and deaths.Results (1)As OD was concerned,there was a statis- tically significant difference in optical density of HMGB-1 on 1st and 3rd days between the two groups of multiple injury patients(t=4.411,P

Objective To investigate the feasibility of ~1H-MR spectroscopy(~1H-MRS)imaging to quantitatively detect fatty liver.Methods Twenty patients with fatty liver and 11 healthy volunteers underwent plain CT scan,conventional MR imaging and ~1H-MRS analysis.The blood lipid and liver function were tested on the same day as the MR examination.~1H-MRS sequence measured the peaks of H_2O and lipid,and the areas under the peaks.The relative contents of the lipid compound were calculated,and compared with the results of CT scan and liver function tests.Results The CT values of the normal group and the fatty liver group were(59?9)HU and(24?11)HU respectively.On ~1H-MRS a protruding high H_2O peak and a flat low lipid peak were observed in the normal group,while the protruding high H_2O peak and a high lipid peak appeared in the fatty liver group.The values of lipid peak in the normal group and the fatty liver group were(0.05?0.01)?10~5,(0.70?0.24)?10~5 respectively(t=4.32,P0.05),the areas under the lipid peak were(1.36?0.73)?10~9、(2.35?1.15)?10~9 respectively(t=5.21,P0.05).Conclusion ~1 H-MRS imaging is feasible to quantitatively detect liver fat and is a non-invasive method for detecting early fatty liver.